Acid sensitivity of neomycin-resistant mutants of Oenococcus oeni: a relationship between reduction of ATPase activity and lack of malolactic activity

Tourdot-Maréchal, Raphaëlle

doi:10.1016/s0378-1097(99)00377-8

Cited by 39 publications

(32 citation statements)

References 22 publications

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“…Similar to mutants in other studies (23,24,52), these mutants had slower growth at pH 4.5 (results not shown). In addition, the ATPase activities of mutant cells were lower than those of the parent strain, and the four mutant strains selected for further study had 52 to 62% less F 0 F 1 -ATPase activity than the parent strain, which corresponded to activities of 2.9 to 3.6 mol P i min Ϫ1 mg Ϫ1 (Fig.…”

Section: Resultssupporting

confidence: 89%

Survival of Probiotic Lactobacilli in Acidic Environments Is Enhanced in the Presence of Metabolizable Sugars

Corcoran

Stanton

Fitzgerald

et al. 2005

Appl Environ Microbiol

441

257

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Lactobacillus rhamnosus GG is an industrially significant probiotic strain with proven health benefits. In this study, the effect of glucose on L. rhamnosus GG survival was analyzed in simulated gastric juice at pH 2.0. It was found that the presence of 19.4 mM glucose resulted in up to 6-log 10 -enhanced survival following 90 min of exposure. Further work with dilute HCl confirmed that glucose was the sole component responsible. Comparative analysis with other Lactobacillus strains revealed that enhanced survival was apparent in all strains, but at different pH values. The presence of glucose at concentrations from 1 to 19.4 mM enhanced L. rhamnosus GG survival from 6.4 to 8 log 10 CFU ml ؊1 in simulated gastric juice. The mechanisms behind the protective effect of glucose were investigated. Addition of N,N-dicyclohexylcarbodiimide to simulated gastric juice caused survival to collapse, which was indicative of a prominent role in inhibition of F 0 F 1 -ATPase. Further work with neomycin-resistant mutants that exhibited 38% to 48% of the F 0 F 1 -ATPase activity of the parent confirmed this, as the survival in the presence of glucose of these mutants decreased 3 ؋ 10 6 -fold compared with the survival of the wild type (which had a viability of 8.02 log 10 CFU ml ؊1 ). L. rhamnosus GG survival in acidic conditions occurred only in the presence of sugars that it could metabolize efficiently. To confirm the involvement of glycolysis in the glucose effect, iodoacetic acid was used to inhibit glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity. The reduction in GAPDH activity caused survival to decrease by 8.30 log 10 CFU ml ؊1 in the presence of glucose. The data indicate that glucose provides ATP to F 0 F 1 -ATPase via glycolysis, enabling proton exclusion and thereby enhancing survival during gastric transit.

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Section: Resultssupporting

confidence: 89%

Survival of Probiotic Lactobacilli in Acidic Environments Is Enhanced in the Presence of Metabolizable Sugars

Corcoran

Stanton

Fitzgerald

et al. 2005

Appl Environ Microbiol

441

257

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“…In our laboratory, we isolated spontaneous ATPase-deficient mutants of O. oeni (26). The acid sensitivity of these mutants suggested a role for H ϩ -ATPase in the acid tolerance of O. oeni, like that demonstrated for other lactic acid bacteria.…”

mentioning

confidence: 56%

“…The parental strain O. oeni IOB84.13 (Oenological Institute of Bordeaux) and neomycin-resistant strains 9.01.07 and 6.27.06 obtained previously (26) were cultured at 30°C and pH 5.3 in FT80 medium modified (FT80m) as described by Labarre et al (13). L. mesenteroides subsp.…”

Section: Methodsmentioning

confidence: 99%

Absence of Malolactic Activity Is a Characteristic of H⁺-ATPase-Deficient Mutants of the Lactic Acid BacteriumOenococcus oeni

Galland

Tourdot-Maréchal

Abraham

et al. 2003

Appl Environ Microbiol

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The lack of malolactic activity in H ؉ -ATPase-deficient mutants of Oenococcus oeni selected previously was analyzed at the molecular level. Western blot experiments revealed a spot at 60 kDa corresponding to the malolactic enzyme only in the parental strain. Moreover, the mleA transcript encoding the malolactic enzyme was not detected by reverse transcription (RT)-PCR analysis of mutants. These results suggest that the malolactic operon was not transcribed in ATPase-deficient mutants. The mleR gene encoding a LysR-type regulatory protein which should be involved in expression of the malolactic genes was described previously for O. oeni. Results obtained in this study show that the mleR transcript was not detected in the mutants by RT-PCR. No mutation in the nucleotide sequences of the mleR gene and the malolactic operon was found. The effect of a reduction in H ؉ -ATPase activity on L-malate metabolism was then investigated by using other malolactic bacteria. Spontaneous H ؉ -ATPase-deficient mutant strains of Lactococcus lactis and Leuconostoc mesenteroides were isolated by using neomycin resistance. Two mutants were selected. These mutants exhibited ATPase activities that were reduced to 54 and 70% of the activities obtained for the L. lactis and L. mesenteroides parental strains, respectively. These mutants were also acid sensitive. However, in contrast to the ATPasedeficient mutants of O. oeni, activation of L-malate metabolism was observed with the L. lactis and L. mesenteroides mutants under optimal or acidic growth conditions. These data support the suggestion that expression of the genes encoding malolactic enzymes in O. oeni is regulated by the mleR product, as it is in L. lactis. Nevertheless, our results strongly suggest that there is a difference between the regulation of expression of the malolactic locus in O. oeni and the regulation of expression of this locus in less acidophilic lactic acid bacteria.

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“…4). This ATP production is probably the basis for the implication of MLF in O. oeni acid tolerance (229). Furthermore, due to the pK difference of the carboxylic groups of malate and lactate, the external replacement of malate by lactate results in alkalinization of the medium.…”

Section: Mechanisms Of Resistancementioning

confidence: 99%

Surviving the Acid Test: Responses of Gram-Positive Bacteria to Low pH

Cotter

Hill

2003

Microbiol Mol Biol Rev

1,042

828

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Gram-positive bacteria possess a myriad of acid resistance systems that can help them to overcome the challenge posed by different acidic environments. In this review the most common mechanisms are described: i.e., the use of proton pumps, the protection or repair of macromolecules, cell membrane changes, production of alkali, induction of pathways by transcriptional regulators, alteration of metabolism, and the role of cell density and cell signaling. We also discuss the reponses of Listeria monocytogenes, Rhodococcus, Mycobacterium, Clostridium perfringens, Staphylococcus aureus, Bacillus cereus, oral streptococci, and lactic acid bacteria to acidic environments and outline ways in which this knowledge has been or may be used to either aid or prevent bacterial survival in low-pH environments

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Acid sensitivity of neomycin-resistant mutants of Oenococcus oeni: a relationship between reduction of ATPase activity and lack of malolactic activity

Cited by 39 publications

References 22 publications

Survival of Probiotic Lactobacilli in Acidic Environments Is Enhanced in the Presence of Metabolizable Sugars

Survival of Probiotic Lactobacilli in Acidic Environments Is Enhanced in the Presence of Metabolizable Sugars

Absence of Malolactic Activity Is a Characteristic of H⁺-ATPase-Deficient Mutants of the Lactic Acid BacteriumOenococcus oeni

Surviving the Acid Test: Responses of Gram-Positive Bacteria to Low pH

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Acid sensitivity of neomycin-resistant mutants of Oenococcus oeni: a relationship between reduction of ATPase activity and lack of malolactic activity

Cited by 39 publications

References 22 publications

Survival of Probiotic Lactobacilli in Acidic Environments Is Enhanced in the Presence of Metabolizable Sugars

Survival of Probiotic Lactobacilli in Acidic Environments Is Enhanced in the Presence of Metabolizable Sugars

Absence of Malolactic Activity Is a Characteristic of H+-ATPase-Deficient Mutants of the Lactic Acid BacteriumOenococcus oeni

Surviving the Acid Test: Responses of Gram-Positive Bacteria to Low pH

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Absence of Malolactic Activity Is a Characteristic of H⁺-ATPase-Deficient Mutants of the Lactic Acid BacteriumOenococcus oeni